CAJ | 학술논문

目的：探讨Grp78基因过表达慢病毒载体的构建、包装和滴度检测。方法采用慢病毒载体，运用基因工程技术，获取目的基因片段并构建重组质粒，然后制备感受态细胞并转化，将重组质粒导入感受态细胞；用PCR技术鉴定阳性克隆和进行DNA序列分析及对慢病毒包装和滴度检测。结果阳性克隆测序比对结果说明测通。熔解曲线中既没出现杂峰也没出现主峰的异常增宽，表明实验中未出现污染、引物二聚体和非特异性扩增。结论Grp78基因过表达慢病毒载体构建和包装成功。
목적：탐토Grp78기인과표체만병독재체적구건、포장화적도검측。방법채용만병독재체，운용기인공정기술，획취목적기인편단병구건중조질립，연후제비감수태세포병전화，장중조질립도입감수태세포；용PCR기술감정양성극륭화진행DNA서렬분석급대만병독포장화적도검측。결과양성극륭측서비대결과설명측통。용해곡선중기몰출현잡봉야몰출현주봉적이상증관，표명실험중미출현오염、인물이취체화비특이성확증。결론Grp78기인과표체만병독재체구건화포장성공。
Objective To constructe ,package and identificate the lentiviral vector with overexpression gene Grp78 .Methods We used lentiviral vector and genetic engineering technology to obtain the aim gene fragment and to constructe recombinant plas‐mid .we prepared competent cells and transform the cells .Through positive clone sequencing ,lentivirus was packaged and virus titer was tested .Results Positive cloning sequence comparison results show that the test was passed .Melt curve did not appear mixed peak ,also did not appear abnormal peak broadening .It means that does not appear pollution ,primer dimers and nonspecific amplifi‐cation in the experiments .Conclusion The construction ,packaging and identification of lentiviral vector with over expression gene Grp78 are sucessful .